Species
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Escherichia coli (Migula) Castellani and Chalmers
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Depositors
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Harvard Univ., KL Strauch, Harvard Univ.
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Applications
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produces protein methyl-accepting chemotaxis protein I (inner membrane protein)
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Vector
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Construct size (kb): 6.0 DESCRIPTION OF VECTOR COMPONENT: Name of vector: pBR322 Intact vector size: 4.363 Type of vector: plasmid Cloning sites: EcoRI ClaI HindIII EcoRV BamHI SphI SalI XmaIII NruI BspMI BsmI StyI AvaI BalI BspMII PvuII Tth111I NdeI AflIII PpaI PstI PvuI ScaI SspI AatII Polylinker sites: Construction: pBR313 Host range: Escherichia coli Features (with orientation and position when available): marker(s): tetR replicon: pMB1 marker(s): ampR Cross references: DNA Seq. Acc.: J01749
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Insert
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DNA: genomic DESCRIPTION OF INSERT COMPONENT: Genome: Escherichia coli Gene symbol: phoA Gene name: alkaline phosphatase Contains complete coding sequence?: U Localization: 9 min Genome: Escherichia coli Gene symbol: tsr Gene name: methyl-accepting chemotaxis protein I (inner membrane protein) Contains complete coding sequence?: U Type of DNA: genomic Insert end: Insert end: Insert size (kb): Cross references: Gene product: methyl-accepting chemotaxis protein I (inner membrane protein) [tsr] Target Gene: methyl-accepting chemotaxis protein I (inner membrane protein)
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Biosafety Level
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1
Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.
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Shipping Information
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Distributed: freeze-dried
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Comments
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Restriction digests of the clone give the following sizes (kb): BamHI--uncut; EcoRI--4.4, 1.55, 0.35; PstI--6.0; HindIII--6.0. If the chimeric protein is degraded by proteases, the alkaline phosphatase becomes soluble in the periplasm and retains activity. The host chromosome contains an insertionally inactivated gene for the periplasmic protease htrA (degP), and thus shows reduced protease activity and increased stability of periplasmic proteins. pKS3 encodes a Tsr-alkaline phosphatase chimeric protein, with Tsr being N-terminal and anchoring the protein in the membrane. The alkaline phosphatase portion is exposed in the periplasm and retains activity.
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Classification
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Enterobacteriaceae, Escherichia
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References
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Beckwith JR, Strauch KL. Periplasmic protease mutants of Escherichia coli. US Patent 4,946,783 dated Aug 7 1990
Strauch KL, Beckwith J. An Escherichia coli mutation preventing degradation of abnormal periplasmic proteins. Proc. Natl. Acad. Sci. USA 85: 1576-1580, 1988. PubMed: 3278319
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Disclosure
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This material is cited in a US or other Patent and may not be used to
infringe the claims. Depending on the wishes of the Depositor, ATCC may be
required to inform the Patent Depositor of the party to which the material was
furnished. This material may not have been produced or characterized by ATCC.
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